Global gene expression profiling of end-stage dilated cardiomyopathy using a human cardiovascular-based cDNA microarray
Global gene expression profiling of end-stage dilated cardiomyopathy using a human cardiovascular-based cDNA microarray
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DOI：
10.1016/s0002-9440(10)61153-4
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发表时间：
2002-06-01
影响因子：
6
通讯作者：
Liew, CC
中科院分区：
文献类型：
--
作者：
Barrans, JD;Allen, PD;Liew, CC
为了获得来自终末期扩张型心肌病（DCM）的心力衰竭的基因组图谱，我们探索了使用非冗余10，848-元素人心血管表达序列标签玻璃载玻片cDNA微阵列的表达分析。从7名接受移植的患者的左心室游离壁和5名非衰竭心脏样本中提取RNA。Cy 3-和Cy 5-标记（和反向染料标记）的cDNA探针从个体患病或非衰竭的成人心脏RNA合成，并与阵列杂交。超过100个转录物在DCM中一致差异表达>1.5倍（与合并的非衰竭心脏相比，P < 0.05）。心钠素在扩张型心肌病患者中表达上调（与非衰竭患者相比，19倍，P < 0.05），同时还有许多肌节和细胞骨架蛋白（如心肌肌钙蛋白、原肌球蛋白）、应激反应蛋白（如HSP 40、HSP 70）和转录/翻译调节因子（如CCAAT盒结合因子、eIF-1AY）。在细胞信号传导通道和介质中观察到的下调最显著，特别是参与Ca 2+途径的那些（Ca 2 +/钙调蛋白依赖性激酶，1，4，5-三磷酸肌醇受体，SERCA）。最有趣的是几个新的，心脏富集表达序列标签的共表达。定量实时逆转录酶-聚合酶链反应的选择这些克隆验证表达。我们的研究使用迄今为止最大的人类心脏特异性cDNA微阵列提供了DCM的初步分子特征。
To obtain a genomic portrait of heart failure derived from end-stage dilated cardiomyopathy (DCM), we explored expression analysis using the CardioChip, a non-redundant 10,848-element human cardiovascular-based expressed sequence tag glass slide cDNA microarray constructed in-house. RNA was extracted from the left ventricular free wall of seven patients undergoing transplantation, and five nonfailing heart samples. Cy3- and Cy5-labeled (and reverse dye-labeled) cDNA probes were synthesized from individual diseased or nonfailing adult heart RNA, and hybridized to the array. More than 100 transcripts were consistently differentially expressed in DCM >1.5-fold (versus pooled nonfailing heart, P < 0.05). Atrial natriuretic peptide was found to be up-regulated in DCM (19-fold compared to nonfailing, P < 0.05), as well as numerous sarcomeric and cytoskeletal proteins (eg, cardiac troponin, tropomyosin), stress response proteins (eg, HSP 40, HSP 70), and transcription/translation regulators (eg, CCAAT box binding factor, eIF-1AY). Down-regulation was most prominently observed with cell-signaling channels and mediators, particularly those involved in Ca2+ pathways (Ca2+/calmodulin-dependent kinase, inositol 1,4,5-trisphosphate receptor, SERCA). Most intriguing was the co-expression of several novel, cardiac-enriched expressed sequence tags. Quantitative real-time reverse transcriptase-polymerase chain reaction of a selection of these clones verified expression. Our study provides a preliminary molecular profile of DCM using the largest human heart-specific cDNA microarray to date.